細(xì)胞STR鑒定簡(jiǎn)介：

細(xì)胞STR鑒定應(yīng)用于生物醫(yī)學(xué)研究領(lǐng)域的哺乳動(dòng)物細(xì)胞被錯(cuò)誤鑒定和交叉污染的問題，一直是一個(gè)普遍存在的突出問題。

據(jù)統(tǒng)計(jì)，國(guó)外實(shí)驗(yàn)室有20%左右的細(xì)胞株被錯(cuò)誤鑒定和交叉污染，NIH和ATCC近兩年都對(duì)此發(fā)出呼吁，要求研究者對(duì)細(xì)胞進(jìn)行鑒定。近年來，大量研究表明STR基因分型方法是進(jìn)行細(xì)胞交叉污染和性質(zhì)鑒定的最有效和準(zhǔn)確的方法之一，STR基因分型應(yīng)用于細(xì)胞鑒定已被ATCC等機(jī)構(gòu)強(qiáng)烈推薦。美國(guó)的ATCC細(xì)胞庫(kù)、德國(guó)的DSMZ細(xì)胞庫(kù)以及日本的JCRB細(xì)胞庫(kù)等為STR分型提供了各細(xì)胞株的數(shù)據(jù)供比對(duì)。

STR（Short Tandem Repeat，短串聯(lián)重復(fù)序列）基因位點(diǎn)由長(zhǎng)度為3-7個(gè)堿基對(duì)的短串連重復(fù)序列組成，這些重復(fù)序列廣泛存在于人類基因組中，可作為高度多態(tài)性標(biāo)記，被稱為細(xì)胞的DNA指紋，其可通過PCR（聚合酶鏈?zhǔn)椒磻?yīng)）來檢測(cè)。STR基因座位上的等位基因可通過擴(kuò)增區(qū)域內(nèi)重復(fù)序列的拷貝數(shù)的不同來區(qū)分，在毛細(xì)管電泳分離之后可通過熒光檢測(cè)來識(shí)別。隨后通過一定的計(jì)算方法，即可根據(jù)所得的STR分型結(jié)果與專業(yè)的細(xì)胞STR數(shù)據(jù)庫(kù)比對(duì)從而推算出樣品所屬的細(xì)胞系或可能的交叉污染的細(xì)胞系名稱。

細(xì)胞STR鑒定服務(wù)流程：

細(xì)胞STR鑒定樣本要求：

細(xì)胞系錯(cuò)誤鑒定所造成的后果：

★ 細(xì)胞系的損失；

★ 時(shí)間和金錢的損失；

★ 在公眾領(lǐng)域（文獻(xiàn)、專利等）提供錯(cuò)誤信息；

★ 造成實(shí)驗(yàn)結(jié)果不一致或不可重復(fù)；

★ ……

何時(shí)需做細(xì)胞STR鑒定？

1、發(fā)表文章或申請(qǐng)課題經(jīng)費(fèi)前；

2、使用細(xì)胞進(jìn)行臨床治療（試驗(yàn)）前；

3、準(zhǔn)備凍存保種或已凍存多年的細(xì)胞；

4、一個(gè)涉及到細(xì)胞試驗(yàn)項(xiàng)目開始/結(jié)束時(shí)；

5、新得到的細(xì)胞或?qū)嶒?yàn)室傳5代以上的細(xì)胞；

6、細(xì)胞系表現(xiàn)不穩(wěn)定或結(jié)果與預(yù)期差別較大。

已開始要求細(xì)胞鑒定的期刊：

★ Nature

★ BioTechniques

★ Cancer Research

★ Cancer Discovery

★ Clinical Cancer Research

★ Molecular Cancer Research

★ Cancer Prevention Research

★ International Journal of Cancer

★ Molecular Cancer Therapeutics

★ Cell Biochemistry and Biophysics

★ Cancer Epidemiology, Biomarkers & Prevention

★ In Vitro Cellular & Developmental Biology - Animal

★ ......

人源細(xì)胞STR位點(diǎn)信息：

為進(jìn)一步提高鑒定的分辨率，除了包含ATCC檢測(cè)的8個(gè)STR和1個(gè)性別決定位點(diǎn)Amelogenin外，另新增了12個(gè)高度多態(tài)性位點(diǎn)，共檢測(cè)21個(gè)STR位點(diǎn)。

參考文獻(xiàn)：

1. Chatterjee, R. (2007) Cell biology. Cases of mistaken identity. Science 315, 928-31.

2. Ruiz Bravo, N. and Gottesman, M. (2007) Notice regarding authentication of cultured cell lines.

3. Yoshino, K. et al. (2006) Essential role for gene profiling analysis in the authentication of human cell lines. Human Cell 19, 43-8.

4. Szibor, R. et al. (2003) Cell line DNA typing in forensic genetics—the necessity of reliable standards. Forensic Sci. Int. 138, 37-43.

5. Dirks, W.G. et al. (2005) Short tandem repeat DNA typing provides an international reference standard for authentication of human cell lines. ALTEX 22, 103-9.

6. Masters, J.R. et al. (2001) Short tandem repeat profiling provides an international reference standard for human cell lines. Proc. Natl. Acad. Sci. USA 98, 8012-7.

7. (2001) Verify cell line identity with DNA profiling. ATCC Connection: Newsletter of The American Type Culture Collection 21, 1-2.

8. Krenke, B. et al. (2002) Validation of a 16-locus fluorescent multiplex system. J. Forensic Sci. 47, 773-85.

9. Levinson, G. and Gutman, G.A. (1987) Slipped-strand mispairing: A major mechanism for DNA sequence evolution. Mol. Biol. Evol. 4, 203-21.

10. Schlotterer, C. and Tautz, D. (1992) Slippage synthesis of simple sequence DNA. Nucleic Acids Res. 20, 211-5.

11. Smith, J.R. et al. (1995) Approach to genotyping errors caused by nontemplated nucleotide addition by Taq DNA polymerase. Genome Res. 5, 312-7.

12. Magnuson, V.L. et al. (1996) Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: Implications for PCR-based genotyping. BioTechniques 21, 700-9.

13. Walsh, P.S., Fildes, N.J. and Reynolds, R. (1996) Sequence analysis and characterization of stutter products at the tetranucleotide repeat locus vWA. Nucleic Acids Res. 24, 2807-12.

14. Moller, A., Meyer, E. and Brinkmann, B. (1994) Different types of structural variation in STRs: HumFES/FPS, HumVWA and HumD21S11. Int. J. Leg. Med. 106, 319-23.

15. Brinkmann, B., Moller A. and Wiegand, P. (1995) Structure of new mutations in 2 STR systems. Int. J. Leg. Med. 107, 201-3.

16. Griffiths, R. et al. (1998) New reference allelic ladders to improve allelic designation in a multiplex STR system. Int. J. Legal Med. 111, 267-72.

17. Bär, W. et al. (1997) DNA recommendations: Further report of the DNA Commission of the ISFH regarding the use of short tandem repeat systems. Int. J. Legal Med. 110, 175-6.

18. Gill, P. et al. (1997) Considerations from the European DNA Profiling Group (EDNAP) concerning STR nomenclature. Forensic Sci. Int. 87, 185-92.